Microorganism evaluation system

ABSTRACT

A microorganism evaluation system comprising a viewing section for image acquisition, the viewing section comprising a viewing port configured to accommodate a fluid flow, at least one independently controlled imaging light source operably installed in the viewing section and configured to selectively illuminate the viewing port, and at least one independently controlled light stimulation device operably installed in the viewing section and configured to selectively emit light for invoking a motile response in a microorganism within the fluid flow in the viewing port, whereby the system synchronizes illumination of the at least one imaging light source and the at least one light stimulation device of the viewing section.

RELATED APPLICATIONS

This is a continuation application and so claims the benefit pursuant to35 U.S.C. § 120 of a prior filed and currently pending U.S. applicationhaving Ser. No. 15/105,280 and filing date of Jun. 16, 2016, andentitled “Microorganism Evaluation System,” which is itself a U.S.national stage entry from an international PCT patent application havingserial number PCT/US2014/070420 and filing date of Dec. 15, 2014, andentitled “Microorganism Evaluation System,” itself claiming prioritypursuant to 35 U.S.C. § 119(e) to and is entitled to the filing date ofa prior U.S. provisional application having Ser. No. 61/916,343 andfiling date of Dec. 16, 2013, and entitled “Microorganism EvaluationSystem Viewing Section.” The contents of the aforementioned applicationsare incorporated herein by reference.

INCORPORATION BY REFERENCE

Applicant hereby incorporates herein by reference any and all patentsand published patent applications cited or referred to in thisapplication.

TECHNICAL FIELD

Aspects of this invention relate generally to evaluation systems, andmore particularly to a microorganism evaluation system and a viewingsection thereof configured for both stimulating and acquiring images ofmicroorganisms within a fluid.

BACKGROUND ART

By way of background, a number of industries are affected by regulationsrelating to water treatment, such as ballast water treatment systems(“BWTS”) on ships and the like. Such regulations require thatmicroorganisms be effectively treated (killed) by the BWTS before suchwater is returned to the ocean or other body of water. Generallyspeaking, Zooplankton in the size range of approximately 10 to 50microns is an “indicator” microorganism used to determine theeffectiveness of treatment, though it will be appreciated that otherorganisms in alternative size ranges are possible depending on thecontext and other factors, such that organisms greater than 50 micronsmay also be the “indicators.” In the art, monitoring of theeffectiveness of such BWTS has largely been handled through samplessubmitted to a lab, there most often involving human examination under amicroscope. Such approaches to compliance assessment have numerousshortcomings in terms of accuracy, speed, and cost. Similarly, flowcytometry systems, though typically offering relatively higherthroughput, are also lacking in terms of viability determination(determinations regarding whether an organism is living) and portabilityfor field or deployed uses. Applicant has already made improvements oversuch prior art systems by developing new and novel evaluation systemsfor determining whether microorganisms are living, such as disclosed inpending international patent application Ser. No. PCT/US13/46334 filedJun. 18, 2013, and U.S. provisional patent application Ser. No.61/661,011 filed Jun. 18, 2012, to which the international applicationclaims priority, both entitled “Microorganism Evaluation System.” Thecontents of the aforementioned applications are incorporated herein byreference.

DISCLOSURE OF INVENTION

Aspects of the present invention teach certain benefits in constructionand use which give rise to the exemplary advantages described below.

The present invention solves the problems described above by providingnew and novel improvements in or relating to the viewing section of suchmicroorganism evaluation systems wherein image data relating toorganisms within a fluid flow is acquired and, in various embodiments,further stimulation of the organisms is provided for purposes oftriggering a motile response of the organisms that is then detected andcaptured by the imaging equipment, as discussed in detail below.

A primary objective inherent in the above described system and method ofuse is to provide advantages not taught by the prior art.

Another objective is to provide a microorganism evaluation systemcomprising a viewing section for image acquisition, the viewing sectioncomprising: a viewing port configured to accommodate a fluid flow from aviewing section body inlet to a viewing section body outlet; at leastone independently controlled imaging light source operably installed inthe viewing section and configured to selectively illuminate the viewingport; and at least one independently controlled light stimulation deviceoperably installed in the viewing section and configured to selectivelyemit light for invoking a motile response in a microorganism within thefluid flow in the viewing port, whereby the system synchronizesillumination of the at least one imaging light source and the at leastone light stimulation device of the viewing section.

Another objective is to provide such a microorganism evaluation systemwherein the at least one imaging light source is installed in theviewing section so as to provide substantially side illumination withinthe viewing port.

Another objective is to provide such a microorganism evaluation systemhaving one or more further alternative organism stimulation mechanismsincorporated on, in, or adjacent to the viewing section.

Another objective is to provide a method of operating the viewingsection of the microorganism evaluation system, comprising the steps of:activating the independently controlled imaging light source operablyinstalled in the viewing section and configured to selectivelyilluminate the viewing port thereof, the viewing port configured toaccommodate a fluid flow; and activating the independently controlledlight stimulation device operably installed in the viewing section andconfigured to selectively emit light for invoking a motile response in amicroorganism within the fluid flow in the viewing port.

It will be appreciated by those skilled in the art that the exactconfiguration of the apparatus may take a number of forms to suitparticular applications without departing from the spirit and scope ofthe present invention. Accordingly, it will be further appreciated thatthe configuration of the apparatus shown and described is exemplary andthat the invention is not so limited.

Other features and advantages of aspects of the present invention willbecome apparent from the following more detailed description, taken inconjunction with the accompanying drawings, which illustrate, by way ofexample, the principles of aspects of the invention.

BRIEF DESCRIPTION OF DRAWINGS

The accompanying drawings illustrate aspects of the present invention.In such drawings:

FIG. 1 is a perspective view of an operative portion of an exemplarymicroorganism evaluation system, in accordance with at least oneembodiment;

FIG. 2 is an exploded perspective view thereof, in accordance with atleast one embodiment;

FIG. 3 is an enlarged partial perspective view of a first exemplaryviewing section thereof, in accordance with at least one embodiment;

FIG. 4 is a partial sectional view of the exemplary viewing section ofFIG. 3 taken from a first perspective;

FIG. 5 is a further partial sectional view of the exemplary viewingsection of FIG. 3 taken from a second perspective;

FIG. 6 is an enlarged partial sectional view of the exemplary viewingsection of FIG. 3;

FIG. 7 is an enlarged partial perspective view of a second exemplaryviewing section thereof, in accordance with at least one embodiment;

FIG. 8 is a partial sectional view of the exemplary viewing section ofFIG. 7 taken from a first perspective;

FIG. 9 is a further partial sectional view of the exemplary viewingsection of FIG. 7 taken from a second perspective;

FIG. 10 is an enlarged partial sectional view of the exemplary viewingsection of FIG. 7;

FIG. 11 is a timing diagram thereof, in accordance with at least oneembodiment;

FIG. 12 is an enlarged partial sectional view of a third exemplaryviewing section thereof, in accordance with at least one embodiment;

FIG. 13 is a further enlarged partial sectional view of the exemplaryviewing section of FIG. 12;

FIG. 14 is a further partial sectional view of the exemplary viewingsection of FIG. 12;

FIG. 15 is an enlarged partial sectional view of a fourth exemplaryviewing section thereof;

FIG. 16 is a further enlarged partial sectional view of the exemplaryviewing section of FIG. 15;

FIG. 17 is an enlarged partial perspective view of a fifth exemplaryviewing section thereof, in accordance with at least one embodiment;

FIG. 18 is a partial sectional view of the exemplary viewing section ofFIG. 17 taken from a first perspective; and

FIG. 19 is an enlarged partial sectional view of the exemplary viewingsection of FIG. 17 taken from a second perspective.

The above described drawing figures illustrate aspects of the inventionin at least one of its exemplary embodiments, which are further definedin detail in the following description. Features, elements, and aspectsof the invention that are referenced by the same numerals in differentfigures represent the same, equivalent, or similar features, elements,or aspects, in accordance with one or more embodiments.

MODES FOR CARRYING OUT THE INVENTION

The above described drawing figures illustrate aspects of the inventionin at least one of its exemplary embodiments, which are further definedin detail in the following description.

As an overview, and with reference to the perspective view of FIG. 1,the exemplary sample acquisition system 20 has four main hardwarecomponents or sections, which are discussed in turn below, with thefocus herein being on being on the third: (1) a microorganismstimulation section 80; (2) a flow normalizing section 100; (3) aviewing section 120; and (4) an outlet section 180. As shown, there maybe one such arrangement or two or more, more about which is said below.There are or may also be related tanks, tubes, filters, pumps, and otheraspects, whether or not shown, that may facilitate the collection andprocessing of a fluid sample according to aspects of the invention,which could be necessary in particular contexts but are neverthelessancillary components that can be substituted for by other equivalentstructure now known or later developed and so are not the focus of thepresent invention. It will be appreciated by those skilled in the artthat the exact configuration of the system and its four main sections orfeatures, again in any number and whether in parallel or in series, maytake a number of forms to suit particular applications without departingfrom the spirit and scope of the present invention. Accordingly, it willbe further appreciated that the configurations of the system shown anddescribed are exemplary and that the invention is not so limited.

Turning now to FIG. 1, there is shown a perspective view of an operativeportion of an exemplary microorganism evaluation system 20 according toaspects of the present invention. Fundamentally, it will be appreciatedthat in the exemplary embodiment of fluid flow sampling, the disclosedevaluation system 20 is directed to or embodies a method by which such afluid flow is first acted on or subjected to some sort of input in orderto stimulate or induce a motile response from living microorganismswithin the flow, and then to visually observe and acquire image datarelative to such a motile response for the purpose of determiningwhether any organisms within the fluid sample are living. The system 20comprises, in the exemplary embodiment, a first or primary, relativelylarger microorganism stimulation section 80′ defining a disorientationspiral 82′ fed by tubing 76′ via coupling 86′ that is itself sampled bya secondary, relatively smaller microorganism stimulation section 80defining a disorientation spiral 82 fed by tubing 76 via coupling 86,such sampling being as by isokinetic sampling, for example. Though notshown in the present application, except for the sectional view of FIG.14 showing the secondary microorganism stimulation section 80 in oneillustrative embodiment, it will be appreciated based on theapplications incorporated by reference herein that within each of thestimulation sections 80, 80′ there may be formed a disorientation spiralor helical flow path configured to induce the fluid in the spiral torotate around the tubular or helical axis, which will stimulate(agitate) the inertial sensing mechanisms found within themicroorganisms. Thus, the previously disclosed inertial stimulationsections 80, 80′ are one example of a means by which to induce a motileresponse within a living organism. Herein are disclosed at least sixother such means or improvements thereto, each of which is itselfexemplary and which it will be appreciated may be employed alone or invarious combinations depending on the context, such that any particularcombination of stimulation means illustrated herein is merely exemplaryof features and aspects of the invention and expressly non-limiting.

With continued reference to FIG. 1 and further with reference to theexploded perspective view of FIG. 2, within the off-line samplingportion of the evaluation system 20, from the smaller inertialstimulation section 80 the flow proceeds to the flow normalizing section100 defining an inlet chute 102, such components being joined viacoupling 90 of the stimulation section 80 and coupling 106 of the flownormalizing section 100. From the flow normalizing section 100, the flowcontinues into the viewing section 120. Similarly, in the main fluidflow, from the larger inertial stimulation section 80′ the flow proceedsto the flow normalizing section 100′ defining an inlet chute 102′, suchcomponents being joined via coupling 90′ of the stimulation section 80′and coupling 106′ of the normalizing section 100′, and from the flownormalizing section 100′ into the viewing section 120′. Much more willbe said about the viewing sections 120, 120′ below, as that is theprimary region of the exemplary microorganism evaluation system 20wherein the various new and novel stimulation means and relatedimprovements are or may be employed as set forth herein. Generally, asshown, each viewing section 120, 120′ comprises in the exemplaryembodiment a viewing section body 122, 122′ having an optical systemmount 130, 130′ and one or more illumination ports 148, 148′, and anopposite back plate 138, 138′ for completing and enclosing the viewingsection 120, 120′ inner space that defines the viewing port 144, 144′ ofeach (FIGS. 4 and 8). As a threshold matter, it is to be understood thatthe illustrated hardware components—here essentially the microorganismstimulation section 80, 80′, the flow normalizing section 100, 100′, andthe viewing section 120, 120′, as well as the outlet section 180, 180′leading away from the viewing section 120, 120′—are merelyrepresentative or illustrative of aspects of the invention and are notlimiting, whether in configuration or arrangement. Simply forillustration regarding scale in the exemplary embodiment, the largerstimulation section 80′ helical flow path may have an inside diameter ofapproximately 13 mm feeding into a viewing section 120′ that isnominally 56.25 mm wide by 12 mm high, as compared to the smallersampling portion in which the stimulation section 80 may have a nominalinside diameter of 2.4 mm and a viewing section 120 that is nominally11.25 mm wide by 3 mm high. The resulting dual system 20 enables morethroughput for use in contexts where larger volumetric or real-timesampling is desired as well as potentially enabling higher accuracy bysecondary line sampling and evaluation within a viewing section 120 thathas a nominal 3 mm depth of field while still allowing an acceptableaggregate throughput by employing the primary line having a nominal 12mm depth of field, or a cross-sectional area of 675 mm² versus the 33.75mm² of the secondary sampling line. It will once again be appreciatedthat such features may be combined in a variety of ways and employ avariety of sizes, shapes, and technologies now known or later developedwithout departing from the spirit and scope of the invention.

Referring now to FIGS. 3-6, there are shown enlarged partial perspectiveand sectional views of the primary, relatively larger line of theexemplary dual sampling microorganism evaluation system 20 generallycomprising the stimulation section 80′, the flow normalizing section100′, the viewing section 120′, and the outlet section 180′. As shownparticularly in FIGS. 4 and 5, in a reverse orientation relative toFIGS. 1-3, the flow normalizing section 100′ generally comprises theinlet chute 102′. It is noted that while a section of tubing 94′ isshown as interconnecting the stimulation section 80′ and the flownormalizing section 100′, or effectively defining he coupling 90′, thisis not necessary, and the two sections 80′, 100′ may instead beconnected directly—it will be appreciated that any such connectivity ofthe respective parts of the system 20 is possible in the presentinvention without departing from its spirit and scope. The chute 102′again has at its proximal end the inlet chute first coupling 106′configured to connect to the stimulation section coupling 90′ andfurther has at is distal end an inlet chute second coupling 108′configured for connecting the inlet chute 102′ to the viewing sectionbody 122′ and the back plate 138′. Once more, though a particular formand geometry of the inlet chute second coupling 108′ is shown, here inthe form of a plate substantially perpendicular to the axis of the inletchute 102′ and having holes formed for the assembly thereof as by boltsor screws to the respective parts of the viewing section 120′, theinvention is not so limited. More notably, it can be seen that theexemplary inlet chute 102′ has an inlet chute body 104′ in which isformed an inlet chute inner bore 110′ along its entire length, whichbore 110′ is substantially tapered or expanding from the entrance to theinlet chute 102′ at the end adjacent the inlet chute first coupling 106′to the exit from the inlet chute 102′ adjacent the inlet chute secondcoupling 108′. Briefly regarding the outlet section 180′ defined by anoutlet chute 182′ that effectively takes the sample flow away from theviewing section 120′ in much the same way, but in reverse, as the inletchute 102′ delivers the sample flow to the viewing section 120′, it canbe seen that the outlet chute inner bore 190′ tapers inwardly orcontracts from the entrance to the outlet chute 182′ at the end adjacentthe outlet chute first coupling 186′ that is connected to the viewingsection body 122′ and back plate 138′ to the exit from the outlet chute182′ adjacent the outlet chute second coupling 188′ that is configuredto connect to other downstream components of the system 20. Again, thoseskilled in the art will appreciate that all such sizes and shapes andconfigurations of such inlet and outlet components, generally directedto slowing the fluid flow as it enters the viewing section 120′ and tospeeding up the fluid flow as it leaves the viewing section 120′, arepossible in the present invention without departing from its spirit andscope.

As best seen in FIG. 4, the exemplary viewing section 120′ is shown withthe viewing section body 122′ upside down and in section along itslength or flow throughput axis. The body 122′ effectively has a viewingsection body inlet 124′ coinciding with the distal end of the inletchute 102′ at its second coupling 108′ and a viewing section body outlet146′ coinciding with the proximal end of the outlet chute 182′ at itsfirst coupling 186′. Three sides of the actual viewing port 144′ or thetrue flow path through the viewing section 120′ are formed by the insidebottom and side surfaces of the back plate 138′ that installs onto theviewing section body 122′ substantially opposite the optical systemmount 130′ and related cavity opening 128′ for allowing viewing into andof the viewing port 144′ by optical equipment (not shown) installed onthe mount 130′. Opposite of or spaced from and substantially parallel tothe back plate 138′ there is positioned a clear or substantiallytransparent viewing plate 136′ (not shown in section or as transparent)offset from the cavity opening 128′, the viewing plate 136′ seatingwithin the viewing section body 122′ so as to form the fourth side ofthe viewing port 144′ through which the fluid sample flows and isvisually inspected and image data relating thereto is acquired asdiscussed in more detail below. With continued reference to FIGS. 3-6,there is also shown multiple illumination ports 148′ intersecting theviewing section body 122′, in each of which there may be installedimaging LEDs or the like so as to illuminate particularly the viewingport 144, more about which will also be said below. Again, any numberand configuration of such illumination ports 148′ and any lighting unitsnow known or later developed may be incorporated into the viewingsection 120′ without departing from the spirit and scope of the presentinvention.

Referring now to FIGS. 7-10, there are shown enlarged partialperspective and sectional views of the secondary, relatively smallerline of the exemplary dual sampling microorganism evaluation system 20that samples from the main line shown in FIGS. 3-6, again generallycomprising the stimulation section 80, the flow normalizing section 100,the viewing section 120, and the outlet section 180, here in a reverseorientation relative to FIGS. 1 and 2, except that the stimulationsection 80 and related feed pipe 76 are in substantially the sameorientation. As shown particularly in FIGS. 8 and 9, the flownormalizing section 100 generally comprises the inlet chute 102 againhaving at its proximal end the inlet chute first coupling 106 configuredto connect to the stimulation section coupling 90 and further having atis distal end an inlet chute second coupling 108 configured forconnecting the inlet chute 102 to the viewing section body 122 and theback plate 138. Once more, though a particular form and geometry of theinlet chute second coupling 108 is shown, here in the form of a platesubstantially perpendicular to the axis of the inlet chute 102 andhaving holes formed for the assembly thereof as by bolts or screws tothe respective parts of the viewing section 120, the invention is not solimited. More notably, it can be seen that the exemplary inlet chute 102has an inlet chute body 104 in which is formed an inlet chute inner bore110 along its entire length, which bore 110 is once again substantiallytapered or expanding from the entrance to the inlet chute 102 at the endadjacent the inlet chute first coupling 106 to the exit from the inletchute 102 adjacent the inlet chute second coupling 108. Brieflyregarding the outlet section 180 defined by an outlet chute 182 thateffectively takes the sample flow away from the viewing section 120 inmuch the same way, but in reverse, as the inlet chute 102 delivers thesample flow to the viewing section 120, it can be seen that the outletchute inner bore 190 tapers inwardly or contracts from the entrance tothe outlet chute 182 at the end adjacent the outlet chute first coupling186 that is connected to the viewing section body 122 and back plate 138to the exit from the outlet chute 182 adjacent the outlet chute secondcoupling 188 that is configured to connect to other downstreamcomponents of the system 20. Again, those skilled in the art willappreciate that all such sizes and shapes and configurations of suchinlet and outlet components, generally directed to slowing the fluidflow as it enters the viewing section 120 and to speeding up the fluidflow as it leaves the viewing section 120, are possible in the presentinvention without departing from its spirit and scope. It is furthernoted in the exemplary embodiment of the viewing section 120 of FIGS.7-10 that the flow path leading into and out of the viewing section 120,and particularly the viewing port 144, curves down and back up again,more about which is said below; such geometrical details are expresslynon-limiting and may simply be a function of spatial constraints,desired depth of field, or other considerations in a particular context.

As best seen in FIG. 8, the exemplary viewing section 120 is shown withthe viewing section body 122 upside down relative to FIGS. 1 and 2 andin section along its length or flow throughput axis. The body 122effectively has a viewing section body inlet 124 coinciding with thedistal end of the inlet chute 102 at its second coupling 108 and aviewing section body outlet 146 coinciding with the proximal end of theoutlet chute 182 at its first coupling 186. Three sides of the actualviewing port 144 or the true flow path through the viewing section 120are formed by the inside bottom and side surfaces of the back plate 138that installs onto the viewing section body 122 substantially oppositethe optical system mount 130 and related cavity opening 128 for allowingviewing into and of the viewing port 144 by optical equipment (notshown) installed on the mount 130. Opposite of or spaced from andsubstantially parallel to the back plate 138 there is positioned a clearor substantially transparent viewing plate 136 (again not shown insection or as transparent) offset from the cavity opening 128, theviewing plate 136 seating within the viewing section body 122 so as toform the fourth side of the viewing port 144 through which the fluidsample flows and is visually inspected and image data relating theretois acquired as discussed in more detail below. With continued referenceto FIGS. 7-10, there is also shown multiple illumination ports 148intersecting the viewing section body 122, in each of which there may beinstalled imaging LEDs or the like so as to illuminate particularly theviewing port 144, more about which will also be said below. Again, anynumber and configuration of such illumination ports 148 and any lightingunits now known or later developed may be incorporated into the viewingsection 120 without departing from the spirit and scope of the presentinvention. For example, and without limitation, in connection with anysuch viewing section 120, 120′ there may be more or less than the fourillumination ports illustrated in the exemplary embodiments, and eachport may be taller or shorter than the ports shown depending on thefocal length of or diffusion from the associated imaging LED or otherlight source in each port as well as other geometrical considerations.Furthermore, in alternative arrangements the illumination ports may beconsiderably reduced in size and incorporated into the side walls of theviewing section as illustrated in the alternative exemplary embodimentof FIGS. 17-19, discussed below.

Turning now to FIG. 11, there is shown a timing diagram relative toessentially the viewing section 120 (FIGS. 1-10), such as thosedisclosed herein or in the prior patent applications incorporated hereinby reference, and particularly to the timing within the viewing section120 of the imager illumination and image acquisition or “shutter” eventsrelative to other lighting or stimulation events, with an objectivebeing to synchronize illumination with image capture to allow forvarious forms of organism stimulation without interference from imagingrequirements. That is, it is desirable that the image illumination andcapture events be effectively synchronized and independently controlledrelative to other stimulation events within the viewing section 120,particularly if those events also involve light, though it will beappreciated by those skilled in the art that in some circumstances suchsynchronization will not be necessary or desirable, such as for examplea scenario wherein there is no light-based stimulation or whereinstimulation light does not interfere with or adversely affectillumination light and image acquisition. In the exemplary smallerviewing section 120, the size of the viewing port 144 as noted above isapproximately 11.25 mm wide by 20 mm long, setting then a viewing portvolume (W×L×H) of approximately 225 mm³ (11.25 mm×20 mm×3 mm) with anominal 3 mm depth of field. Though system flow rates can vary widelybased on a number of factors, the time for the flow, or a particularorganism, to pass through the viewing section 120 is approximately onthe order of one to ten (1-10) seconds. What this translates to, if asubstantial number of image captures of the same organism are to beobtained (e.g., on the order of 25-250 or greater discrete images) onwhich computational analysis is to be performed in determiningorganism-generated movement and thus life, is a cycle time ofapproximately 33 ms based on 30 FPS (frames per second), or moregenerally in the range of approximately fifteen to forty-fivemilliseconds (15-45 ms). As shown in the timing diagram, then, within arepresentative viewing section 120 and based on current imagingtechnology and equipment and the exemplary throughput and othercharacteristics noted, what is represented is the timing of variousevents relative to each other within the exemplary 30 FPS cycle. Thoseskilled in the art will appreciate that the “frames per second”capability of the imaging equipment and thus the cycle time, the flowrate, the geometry of the viewing section 120, and a number of othersuch factors all may be changed to suit particular applications and toaccommodate any related technologies now known or later developed in theart, such that the timing diagram is to be understood as merelyillustrative of features and aspects of the present invention andnon-limiting. That is, it will be appreciated that as imaging technologyimproves and 60, 120, or more FPS image acquisition is achievable, thedwell times within the viewing section 120 can be reduced accordinglywhile still being able to obtain the desired number of discrete imagesor data points per organism moving with the flow through the viewingsection. Relatedly, variables such as the ability and timing to turn onand off, or strobe, the illumination lighting, as discussed furtherbelow, and the typical stimulation response times of the organisms beingevaluated or used as the “indicators” are also factors in establishingthe throughput of the viewing section 120 relative to the data to becaptured. Accordingly, the evaluation system and particularly theviewing section design according to aspects of the present invention canbe “tuned” as desired to suit particular applications—where relativelygreater accuracy and/or lower cost are a priority or the organisms arerelatively slower moving or responding, longer dwell times may bedesired and obtained, even beyond the exemplary 10 seconds, by makingthe viewing section relatively larger or simply slowing down the flowrate through the system, for example, in which case relatively lower FPSimaging equipment and/or relatively slower toggling illumination lightsources may be employed at reduced cost. Or, if relatively higherthroughput is desired instead of or in addition to accuracy or is simplypossible due to relatively fast indicator organism response rates,relatively higher FPS imaging equipment can be used and/or relativelysmaller viewing section geometry, such as in cases where the “package”size is also an important factor. Those skilled in the art willultimately appreciate that a number of such variables are interrelatedand thus that a wide variety of system configurations are possibleaccording to aspects of the present invention without departing from itsspirit and scope. Moreover, it will be appreciated that even theexemplary diagram based on 30 FPS contains certain assumptions and “bestguesses” about the performance or capability of some of the equipment,such that the diagram is not to be taken literally or “to scale,” but isinstead intended to simply convey the overall concept of synchronizingthe illumination and imaging events and having those events beindependently controlled relative to other events within the cycle as afluid flow and hence microorganisms move through the viewing section120. Accordingly, referring first to event “1” on the diagram of FIG.11, there is simply shown a plot representing two cycles, each cyclebeing 33 ms at 30 FPS. Event #2 corresponds to initialization of theimager, which is indicated here as approximately 1.5 ms, starting fromthe beginning of the cycle. Event #3 is the imager illumination event,when whatever light(s) that are to illuminate the viewing port 144 areturned “on” so as to emit for a relatively brief period, hereillustrated as approximately 3 ms or about ten percent (10%) or in therange of five to fifteen percent (5-15%) of the cycle, essentially broadspectrum or broadband light similar to bright sunlight (i.e., a “flash”)so as to properly expose the image sensor. At event #4, substantiallycontemporaneous with the imager illumination event, is the imageacquisition or “shutter” event, here indicated as approximately 2ms—that is, the shutter event would begin just after and end just beforethe imager illumination event. It is also possible, depending on imagingsystem peculiarities, that this relationship could be reverse where theillumination event is shorter than the “shutter” event. After the imageis acquired at event #4, event #5 represents the image data read or datadownload to the image processor or the like (not shown), which may takefrom 5-12 ms or more, depending on a number of factors, starting withthe amount of data acquired and many other factors beyond the scope ofthe present invention (in FIG. 11 event #5 is represented asapproximately 5 ms). Preferably, the image data read event is as shortas possible simply to allow as much time for the data processing event(not shown on diagram) before the next cycle begins and a new set ofimage data is acquired. At event #6, more about which is said below,which runs from essentially the end of the image acquisition event #4 tothe start of the next cycle, microorganisms in the fluid flowsubstantially within the viewing section 120 may be subject to one ormore forms of stimulation during this time, here represented as anexemplary “wavelength stimulator” or stimulation from a particularspectrum of light energy, whether continuous or pulsed. Notably, suchstimulation would have an effective “duty cycle” of approximately eightyto ninety percent (80-90%), or would represent approximately 26-29 ms ofthe nominal 33 ms exemplary cycle. During this period the organisms mayexperience “calm darkness” or other forms of stimulation other thanlight, or may be exposed to light stimulation distinct from therelatively instantaneous imager illumination light source having aduration of approximately 3 ms, which short duration event (event #3)the organisms would likely not respond to. But in terms of lightstimulation during event #6 shown in the timing diagram of FIG. 11, forexample, the stimulation means may be particular wavelengths of lightsuch as a narrow color spectrum light. The source of such light may besimilar LEDs as provided within the illumination ports 148 emitting asingle or different wavelength(s), significantly different LEDs alsopositioned within the same illumination ports 148, or different LEDspositioned elsewhere within the viewing section 120, as will beappreciated particularly with respect to the alternative embodimentsshown in FIGS. 15 and 16 and in FIGS. 17-19, discussed further below.Again, an important aspect of this “illumination synchronization” isthat the illumination for the imager be independently controlledrelative to any other illumination, which not only again provides thebenefit of a relatively substantial “duty cycle” for such stimulation,as noted above, and thus the improved inducement of motion, but alsoimproved detection of motion by isolating the imaging illumination andacquisition events effectively from everything else. That is, anycolored light turned “on” during event #6 could be “off” during theimager events #s 2-4; however, it will again be appreciated that basedon the location of any stimulation light source and the color(wavelength) and intensity of that light, it may simply be left onconstantly as not interfering with or in any way adversely affecting theimaging illumination. Alternatively, irrespective of the illuminationstimulation duty cycle and the type and capability of the lightingunits, any such stimulation light source(s) may be strobed at aparticular frequency rather than left on during all or part of the cyclein order to enhance stimulation for particular organisms. Those skilledin the art will further appreciate that a related benefit of suchillumination synchronization capabilities within the system is thatphotosynthesis may be both caused and detected as a byproduct of suchindependent lighting control. Those skilled in the art will appreciatethat the photosynthesis process in plants absorbs particular spectrum(s)of light energy. The standard detection process is to emit thespectrum(s) of light absorbed and detect the reduction of this energythat is reflected back. According to aspects of the present invention,independent control and synchronization of the imaging, stimulation, andplant life illumination can enhance the optimization of the measurementof chlorophyll levels (photosynthesis) and the other imaging relatedelements, providing yet another valuable data set in the sampleanalysis. For example, plants tend to absorb the non-green spectrums,while zooplankton (animals) tend to be attracted to the green spectrumbecause this is their “food”; thus, photosynthesis might be betterdetected with a blue emitter while zooplankton might be better attractedto a green emitter. With independent lighting control and the potentialfor multiple light sources and varied illumination durations, all such“stimulation” is possible.

By way of further example, and with continued reference to FIG. 11,another exemplary stimulation is pheromone stimulation, or theintermittent emission of pheromones into the fluid flow from a source(not shown) somewhere in or about the viewing section 120. That is, theidea is to provide controlled emission of micro-liters of semiochemicals(bio-signals) into the viewing port 144 in order to excite a motileresponse from an organism such as zooplankton. Such pheromones may beany natural or synthetic products now known or later developed ordiscovered that can be shown to induce a motile response in arepresentative microorganism. In one exemplary embodiment, the pheromonemicro-emitter would be positioned substantially in or near the viewingsection body inlet 124 (FIG. 8) so as to attract a microorganism as itdrifts through the viewing port 144 and goes out the outlet chute 182.As shown in connection with event #7, it is contemplated that thepheromone emission will be at some interval less than once percycle—perhaps every 8th cycle (3 Hz), for example. The emission durationmight be relatively short, generating “drops” of stimulant. Per thediagram, once emitted, the pheromones will “drift” through one or morecycles within the viewing section 120, such as over a period of 50 to100 ms or longer, as compared with the exemplary approximately 33 mscycle, but the actual, discrete pheromone emission event will berelatively less frequent and substantially instantaneous. Notably, thepheromone emission will not be initiated during imaging event #s 2-4,again, so as to not in any way interfere with or compromise the imageacquisition. It will of course be appreciated that any such pheromonestimulation or other such non-illumination stimulation is entirelyoptional but may in certain contexts provide a complimentary source ofstimulation. Finally in terms of the timing diagram of FIG. 11, event#8, shown on the same graph with event #3, again effectively representsthe periods of time between imaging events #s 2-4 when an organism cansimply “swim” undisturbed or be subjected to other forms of sensory orother stimulation. By way of further example regarding the kinds ofstimulation to which organisms may be subjected substantially within theviewing section 120, localized relatively low-energy vibrationstimulation or possibly acoustic stimulation are also contemplated.Either such device (see the exemplary vibratory stimulation device 250of FIGS. 12-16) may again be located substantially near or in theentrance to the viewing port 144, such as in the viewing section bodyinlet 124 (FIG. 8), so as to excite or induce a “flight” response in anorganism such as zooplankton, which will encourage such an organism to“flee” and “escape” with the flow through and out of the viewing section120. The viewing section body outlet 146 and the outlet chute 182 itselfwill seem an “inviting” place for the organisms to retreat to. In thecase of vibration stimulation, it may be located at or near the entranceof the proximity stimulator 240 (discussed below in connection with thealternative embodiment of FIGS. 12-16), and any such device wouldpreferably be mechanically isolated from the rest of the viewing section120. The idea is that this disturbance would not significantly propagatemuch beyond the location of emission. Similarly, some form of acousticstimulation designed to excite the same “flight” response as thevibration stimulator may again be positioned substantially at or nearthe entrance to the viewing section 120. Such an acoustic stimulatorwould be based on acoustic tuning to provide a sense of appropriatedistance and disturbance so as to again encourage the organisms to movetowards the outlet chute 182. And as with the vibration stimulation, theacoustic stimulation device would also preferably be mechanicallyisolated from the rest of the viewing section 120 in order to optimizelocalization and effectiveness.

Referring now to FIGS. 12-16, there are shown enlarged partial sectionalviews of the secondary, relatively smaller line of the exemplary dualsampling microorganism evaluation system 20 that samples from the mainline shown in FIGS. 3-6, analogous to that of FIGS. 7-10 and so againgenerally comprising the stimulation section 80, the flow normalizingsection 100, the viewing section 120, and the outlet section 180, onlynow including as alternative embodiments one or more further exemplarystimulation devices incorporated within the viewing section 120 thereof.First, as shown in FIGS. 12 and 13, there may be positioned within theviewing section body inlet 124 a proximity stimulation device 240, hereshown as what is effectively a plug formed with numerous substantiallyparallel through-holes or sub-chutes 242 effectively communicatingbetween the inlet 124 and the viewing port 144 within the viewingsection 120. The idea is that the zooplankton or other organisms willsense the close proximity of the tunnel walls, or will feel a bit“claustrophobic” within the relatively smaller openings of thesub-chutes 242 and will provide or exhibit a motile response whenentering the “tranquil” and “wide open” waters of the viewing port 144.It will be appreciated by those skilled in the art that a virtuallyinfinite variety of shapes, sizes and configurations of any suchproximity stimulation device 240, and its location, are possible withoutdeparting from the spirit and scope of the present invention. As alsoshown in FIGS. 12 and 13, a vibratory stimulation device 250 may also bepositioned in or adjacent to or integrated within the viewing sectionbody inlet 124 to further stimulate organisms passing into and throughthe inlet 124, with or without the proximity stimulation device 240; butwhere a proximity stimulation device 240 is employed, as shown, it wouldbe preferable to locate such a vibratory stimulation device 250, anacoustic stimulation device, or other such stimulation device closer tothe entrance of the inlet 124 or of the device 240 itself so as to againelicit the “flight” response of the organisms and encourage them to rushdown through the chute(s) and into the “calmer waters” of the viewingsection viewing port 144, where their motile responses would beobserved. Briefly, as shown in FIG. 14, the proximity stimulation device240 may be employed within the viewing section body inlet 124 alone,without the vibratory stimulation device 250 (FIGS. 12 and 13); again,any such combination of stimulation means and mechanisms is possible inthe present invention without departing from its spirit and scope.Furthermore, as shown in FIG. 14, the basic upstream microorganismstimulation section 80 is shown in section as well, revealing portionsof the one or more disorientation spiral loops 84, 88 formed therein.

Turning to FIGS. 15 and 16, there are shown enlarged partial sectionalviews of particularly the viewing section 120 now including a stillfurther exemplary stimulation device in the form of one or more lightsources 260 positioned here within or substantially near the exit of theviewing section body outlet 146 so as to draw or attract organismsthereto. Particularly, there is shown in the exemplary embodiment twooffset light stimulation devices 260 substantially at the exit of theviewing section body outlet 146 or the entrance to the outlet chuteinner bore 190, spaced on either side thereof and so positioned as todraw organisms in the viewing port 144 up and out of the viewing section120 through the outlet 146. The idea is to essentially place a narrowspectrum emitter of some kind in or adjacent the outlet chute 146 of theviewing section 120 that is giving off light that the organisms like andwill swim towards. In that regard, it has been discovered thatzooplankton, for example, has light receptors that would appear to beoptimized for performance relative to the color of their food(phytoplankton) and the light filtering characteristics of water. It isof significant note that wavelengths of approximately 530 nm (green)appear to be effective in attracting zooplankton particularly, thoughother wavelengths such as approximately 475 nm (blue) may also beeffective, such that the invention is expressly not limited to aparticular wavelength or color of light in this context; it will beappreciated that different organisms may be attracted to or preferdifferent colors of light. Any such light stimulation will generally bemost effective when there are not other competing wavelengths inproximity, which again relates back to the above illuminationsynchronization discussion and the idea that not only is there to be nolight interference with the light stimulation devices 260, but there isalso to be no interference by such devices 260 with the imager events #s2-4 in FIG. 11; as such, again, there is benefit in having the imagerlight source(s) and the stimulation light source(s) independentlycontrolled as herein disclosed. Back to the placement of the lightstimulation devices 260 substantially at the exit of the viewing sectionbody outlet 146, or “up the ramp,” it will be appreciated that suchplacement draws the organisms further up the exit and allows foremission of relatively soft light cascading or filtering down into theviewing port 144 even when “on” during the non-image acquisition phaseof the cycle, thereby also rendering less likely any light “pollution”within the viewing port 144 during image acquisition in the event thestimulation or attraction lights 260 were even to be “on”. Relatedly, asshown, the exemplary light stimulation devices 260 have domed,semi-spherical lenses 262 that more readily diffuse light from the LEDor other light source therebeneath (not shown). In other contexts a“point” light versus a diffused light might be preferable. Each suchlight source and lens 262 is shown as being mounted on a base 264,though it will be appreciated that each such light stimulation device260 may be integrated into or mounted in the viewing section body 122 orthe outlet chute 182 in virtually any manner now known or laterdeveloped for securing and positioning such light stimulation devices260 as desired. Again, it will be further appreciated that a variety ofnumber, configuration and location of such devices 260 are possiblewithout departing from the spirit and scope of the invention. By way offurther example and illustration, the inlet and outlet chutes 102 may bedifferent geometries in order to support “line of sight” to the emitteronce the zooplankton or other organisms enter the viewing chamber 144.Or, any such devices 260 may simply be placed nearer to or even in theexit area 146 of the viewing section 120 instead (see, for example, thealternative embodiment shown in FIG. 19 discussed immediately below).

Finally, referring now to FIGS. 17-19, there is shown yet anotherexemplary embodiment of a microorganism evaluation system 20 accordingto further aspects of the present invention, particularly once more inconnection with the viewing section 120. As can be first observed fromthe perspective view of FIG. 17, taken from a vantage point lookingsomewhat down at the optical system mount 130 (more like FIGS. 1-3versus FIGS. 4-6), the inlet section 100 and outlet section 180 areoriented essentially perpendicular to the viewing section 120 ratherthan substantially parallel as in the other illustrated embodiments. Itwill again be appreciated generally that any and all such orientationsare possible within the present invention without departing from itsspirit and scope. Here, it will be appreciated that such a change inorientation of the inlet and outlet sections 100, 180 may be for reasonsof flow considerations or simply spatial or “packaging” constraints. Inany case, as best seen in FIG. 18, the exit of the inlet chute 102 andparticularly its inner bore 110 in the vicinity of the distal coupling108 is yet configured to substantially seamlessly and sealingly engage,join, or otherwise flow into the viewing section body inlet 124 when thecomponents are installed together; a similar arrangement would be trueof the outlet chute 182. As also shown in FIG. 18, once again a clear orsubstantially transparent viewing plate 136 is positioned within theviewing section 120 assembly so as to effectively form one of the foursides of the conduit or flow path defining the viewing port 144 (FIG.19). Here, the viewing plate 136 is configured to seat or be mounted andsecured between the viewing section body 122 and the back plate 138.Though not shown, it will be appreciated from the foregoing that aproximity stimulator 240 and/or vibratory stimulation device 250 (FIG.13) or other such device may be placed within the viewing section bodyinlet 124 as well. Other features and aspects as shown in otherexemplary embodiments herein or as otherwise consistent with orcontemplated by the present disclosure may thus be incorporated withinthe viewing section 120 or other components of the system 20. Turning toFIG. 19, an enlarged sectional view of the viewing section 120 takenmore from the side (and with the viewing plate 136 removed forsimplicity), there is best shown the interior features of the exemplarydevice, and particularly the region of the viewing port 144. Notably,there are shown a series of imaging light sources 150 operably installedin a lengthwise side wall 140 here of the back plate 138 though moregenerally within the viewing section 120 and more particularly theviewing port 144. Though not shown, it will be appreciated that asimilar bank of imaging light sources 150 could also be provided on theopposite side of the viewing port 144 for illumination from both sides.In any case, such imaging light sources 150 may be LEDs or any othersuch technology as now known or later developed in the art, here shownas being contained within the viewing section 120, or incorporated orinstalled within a wall thereof, such that no separate illuminationports 148 (FIG. 7) in which would be installed illumination LEDs areshown or required, though those skilled in the art will appreciate thatsuch light sources at an angle may still be employed in addition to theside-mounted imaging LEDs 150 shown in FIG. 19. Fundamentally, sideillumination light source(s) as with the imaging LEDs 150 may have anumber of advantages in terms of image acquisition and quality. First,such an installation and method mitigates the development of shadowsfrom microorganisms moving about in the viewing port 144, which shadowswould tend to be cancelled out by the opposing illumination emitters150. The arrangement also potentially increases the imaging contrastdeveloped by the microorganisms within the flow in the viewing port 144,as the surface on the body of the microorganism that is closest to theimager will tend to be darker than the side of the microorganism that isilluminated, since the imaging equipment (not shown) is mounted on theoptical system mount 130 (FIGS. 17 and 18) well above the plane of theimaging LEDs 150. Relatedly, the side-mounted LEDs 150 also mitigate thepotential for light energy to be reflected into the imager, includingreflections from the various surfaces found within the viewing section'scavity opening 128, or the space between the viewing port 144 and theimager (not shown) that would be mounted above. In fact, by having theimager separated from the viewing port 144 and emitters 150 by the clearviewing plate 136 (FIG. 18), light will not bounce off of the glass orsimilar material because the illumination is contained in the fluidunderneath the viewing plate 136. With further reference to FIG. 19,each imaging light source 150 is shown somewhat nondescriptly as arectangular configuration LED flush mounted with the side wall 140. Itwill be appreciated by those skilled in the art that any shape or lensconfiguration or angle or again more generally any illuminationtechnology now known or later developed may be employed. It is herecontemplated that the one or more side-mounted imaging LEDs 150 wouldhave relatively wide angle lenses so as to enhance the emission ofrelatively more uniform spatial distribution of light energy, whichshould generate a relatively “flat” and uniform background, as furtheraided by the inherent diffusing nature of the fluid itself within theviewing port 144. While the bank of imaging LEDs 150 on a particularside of the viewing section 120 may be spaced uniformly, as shown inFIG. 19, by design, the imaging LEDs 150 are closer together at themarginal edges of the viewing port 144 or at the respective left andright or inlet and outlet edges of the side wall 140. Toward the centerof the viewing port 144 it would tend to be brightest, where the lightfrom effectively more imaging LEDs 150 would be compounded oraggregated, while toward the marginal edges where there are relativelyfewer LEDs 150 to contribute to the illumination, it would tend to berelatively darker. Thus, by having a higher density or concentration,via closer spacing, of light sources at the margins, the net effect issubstantially uniform or even lighting of the entire viewing port 144.The specific sizes and spacing of the imaging LEDs 150 and their numberper side (nine shown in FIG. 19), are not to be taken literally or toscale or to be in any way limiting, it being appreciated that FIG. 19and the related discussion is merely illustrative of features andaspects of the invention. Accordingly, a wide variety of otherconfigurations and arrangements of imaging light sources, again, whetherfor side or angled or direct illumination or any combination thereof,are to be understood as within the spirit and scope of the invention. Asalso shown in FIG. 19, at least one stimulation light source 260 havinga lens 262 may be positioned substantially at the exit of the viewingport 144 or of the viewing section 120 more generally so as to attractor stimulate microorganisms within the flow as above-described. As alsodescribed herein, the operation of such attractant light source 260 maybe synchronized with any imaging light source 150 in the viewing section120 depending on a variety of factors. Those skilled in the art willagain appreciate that a variety of lighting arrangements and control arepossible without departing from the spirit and scope of the presentinvention.

Aspects of the present specification may also be described as follows:

1. A microorganism evaluation system comprising a viewing section forimage acquisition, the viewing section comprising: a viewing portconfigured to accommodate a fluid flow from a viewing section body inletto a viewing section body outlet; at least one independently controlledimaging light source operably installed in the viewing section andconfigured to selectively illuminate the viewing port; and at least oneindependently controlled light stimulation device operably installed inthe viewing section and configured to selectively emit light forinvoking a motile response in a microorganism within the fluid flow inthe viewing port, whereby the system synchronizes illumination of the atleast one imaging light source and the at least one light stimulationdevice of the viewing section.

2. The system of embodiment 1 wherein the viewing section furthercomprises: a viewing section body; and a back plate, the viewing sectionbody and the back plate together defining the viewing port.

3. The system of embodiment 2 wherein: the viewing section body isformed with at least one illumination port configured to opticallycommunicate with the viewing port; and the at least one imaging lightsource is located in the at least one illumination port.

4. The system of embodiment 2 wherein the at least one imaging lightsource is installed in the viewing section body.

5. The system of embodiment 2 wherein the at least one imaging lightsource is installed in the back plate.

6. The system of embodiment 2 wherein a viewing plate is installedwithin the viewing section substantially opposite the back plate,between the back plate and an optical system cavity opening formed inthe viewing section body and configured to optically communicate withthe viewing port, the viewing plate being substantially transparent; andthe at least one imaging light source is installed in the viewingsection so as to be bounded by the back plate and the viewing plate andthus positioned within the viewing port.

7. The system of embodiment 1 wherein the at least one imaging lightsource is installed in the viewing section so as to providesubstantially side illumination within the viewing port.

8. The system of embodiment 7 wherein a plurality of imaging lightsources are aligned as an array within the viewing port.

9. The system of embodiment 8 wherein the plurality of imaging lightsources are unequally spaced, with a higher density of imaging lightsources at the margins of the viewing port.

10. The system of embodiment 8 wherein the plurality of imaging lightsources are LEDs.

11. The system of embodiment 1 wherein the at least one lightstimulation device is operably installed in the viewing section bodyoutlet.

12. The system of embodiment 1 wherein: an outlet chute is installed onthe viewing section so as to be in fluid communication with the viewingport by way of the viewing section body outlet; and the at least onelight stimulation device is operably installed in the outlet chute.

13. The system of embodiment 1 wherein the at least one lightstimulation device is configured to emit light having a wavelength ofapproximately 530 nm (green).

14. The system of embodiment 1 wherein the at least one lightstimulation device is operated continuously.

15. The system of embodiment 1 wherein the at least one lightstimulation device is pulsed.

16. The system of embodiment 1 wherein the at least one imaging lightsource and the at least one light stimulation device are co-located.

17. The system of embodiment 1 wherein the viewing section furthercomprises a vibratory stimulation device.

18. The system of embodiment 17 wherein the vibratory stimulation deviceis operably installed in the viewing section body inlet.

19. The system of embodiment 17 wherein: an inlet chute is installed onthe viewing section so as to be in fluid communication with the viewingport by way of the viewing section body inlet; and the vibratorystimulation device is operably installed in the inlet chute.

20. The system of embodiment 1 wherein the viewing section furthercomprises a proximity stimulation device.

21. The system of embodiment 20 wherein the proximity stimulation deviceis configured as a plug formed with a plurality of substantiallyparallel sub-chutes.

22. The system of embodiment 21 wherein: an inlet chute is installed onthe viewing section so as to be in fluid communication with the viewingport by way of the viewing section body inlet; and the proximitystimulation device is operably installed in the viewing section bodyinlet so as to be in fluid communication between the inlet chute and theviewing port within the viewing section by way of the pluralitysub-chutes.

23. The system of embodiment 21 wherein: an inlet chute is installed onthe viewing section so as to be in fluid communication with the viewingport by way of the viewing section body inlet; and the proximitystimulation device is operably installed in the inlet chute so as to bein fluid communication with the viewing section body inlet by way of theplurality sub-chutes.

24. The system of embodiment 1 further comprising a microorganismstimulation mechanism selected from the group consisting of acousticenergy and pheromones.

25. The system of embodiment 1 wherein: a cycle of the system is definedas the time period from the start of one discrete illumination event ofthe at least one imaging light source to the next; and each imaginglight source illumination event represents approximately five to fifteenpercent (5-15%) of the cycle.

26. The system of embodiment 25 wherein the cycle is approximatelyfifteen to forty-five milliseconds (15-45 ms).

27. The system of embodiment 25 wherein the illumination event of the atleast one light stimulation device represents approximately eighty toninety percent (80-90%) of the cycle.

28. The system of embodiment 25 wherein the illumination event of the atleast one light stimulation device represents approximately eighty toone-hundred percent (80-100%) of the cycle.

29. The system of embodiment 1 wherein: a first illumination event isassociated with operation of the at least one imaging light source; asecond illumination event is associated with operation of the at leastone light stimulation device; and the first and second events aresubstantially non-overlapping.

30. A microorganism evaluation system comprising a viewing section forimage acquisition, the viewing section comprising: a viewing port invisual communication with an optical system cavity opening formed in theviewing section, the viewing port configured to accommodate a fluidflow; a plurality of imaging light sources operably installed within theviewing port so as to provide substantially side illumination therein;and at least one independently controlled light stimulation deviceoperably installed in the viewing section and configured to selectivelyemit light for invoking a motile response in a microorganism within thefluid flow in the viewing port, whereby the system synchronizesillumination of the plurality of imaging light sources and the at leastone light stimulation device of the viewing section.

31. A method of operating a viewing section of a microorganismevaluation system, comprising the steps of: activating an independentlycontrolled imaging light source operably installed in the viewingsection and configured to selectively illuminate a viewing port thereof,the viewing port configured to accommodate a fluid flow; and activatingan independently controlled light stimulation device operably installedin the viewing section and configured to selectively emit light forinvoking a motile response in a microorganism within the fluid flow inthe viewing port.

32. The method of embodiment 31 wherein prior to the step of activatingthe independently controlled light stimulation device, the methodcomprises the further step of deactivating the imaging light source.

33. The method of embodiment 32 wherein the time the imaging lightsource is activated represents approximately five to fifteen percent(5-15%) of the total time from one imaging light source activation eventto the next, which total time defines a cycle of the system.

34. The method of embodiment 33 comprising the further step ofdeactivating the light stimulation device prior to activating theimaging light source a subsequent time, wherein no imaging light sourceactivation event and light stimulation device activation event overlap.

35. The method of embodiment 34 wherein the light stimulation deviceactivation event represents approximately eighty to ninety percent(80-90%) of the cycle.

36. The method of embodiment 31 wherein the light stimulation deviceactivation event represents approximately eighty to one-hundred percent(80-100%) of a cycle of the system defined as the total time from oneimaging light source activation event to the next.

37. The method of embodiment 31 wherein the steps of activating theindependently controlled imaging light source and activating theindependently controlled light stimulation device are synchronized.

38. The method of embodiment 37 wherein the steps of activating theindependently controlled imaging light source and activating theindependently controlled light stimulation device overlap.

39. The method of embodiment 31 wherein the step of activating theindependently controlled light stimulation device entails emission ofnarrow spectrum light.

40. The method of embodiment 32 wherein the step of emitting narrowspectrum light further entails pulsing the light stimulation device.

41. The method of embodiment 31 comprising the further step of operatingan image acquisition shutter at least partially during the imaging lightsource activation event.

42. The method of embodiment 41 wherein the step of operating the imageacquisition shutter occurs entirely during the imaging light sourceactivation event.

43. The method of embodiment 31 comprising the further step of emittingdrops of pheromones into the fluid flow.

44. The method of embodiment 31 comprising the further step ofintroducing acoustic energy into the fluid flow.

45. The method of embodiment 31 comprising the further step ofintroducing vibrations into the fluid flow.

46. The method of embodiment 31 comprising the further step of passingthe fluid flow through a proximity stimulation device having a pluralityof substantially parallel sub-chutes.

To summarize, regarding the exemplary embodiments of the presentinvention as shown and described herein, it will be appreciated that amicroorganism evaluation system, particularly a viewing section thereof,is disclosed and configured for both stimulating and acquiring images ofmicroorganisms within a fluid, including aspects related tosynchronizing the various illumination events within the system. Becausethe principles of the invention may be practiced in a number ofconfigurations beyond those shown and described, it is to be understoodthat the invention is not in any way limited by the exemplaryembodiments, but is generally able to take numerous forms in doing sowithout departing from the spirit and scope of the invention.

In closing, it is to be understood that although aspects of the presentspecification are highlighted by referring to specific embodiments, oneskilled in the art will readily appreciate that these disclosedembodiments are only illustrative of the principles of the subjectmatter disclosed herein. Therefore, it should be expressly understoodthat the disclosed subject matter is in no way limited to a particularapparatus, methodology, configuration, size, shape, material ofconstruction, protocol, etc., described herein, but may include any suchtechnology now known or later developed without departing from thespirit and scope of the specification. Furthermore, the various featuresof each of the above-described embodiments may be combined in anylogical manner and are intended to be included within the scope of thepresent invention. As such, various modifications or changes to oralternative configurations of the disclosed subject matter can be madein accordance with the teachings herein without departing from thespirit and scope of the present specification. Lastly, the terminologyused herein is for the purpose of describing particular embodimentsonly, and is not intended to limit the scope of the present invention.Accordingly, the present invention is not limited to that precisely asshown and described.

Certain embodiments of the present invention are described herein,including the best mode known to the inventors for carrying out theinvention. Of course, variations on these described embodiments willbecome apparent to those of ordinary skill in the art upon reading theforegoing description. The inventor expects skilled artisans to employsuch variations as appropriate, and the inventors intend for the presentinvention to be practiced otherwise than specifically described herein.Accordingly, this invention includes all modifications and equivalentsof the subject matter recited in the claims appended hereto as permittedby applicable law. Moreover, any combination of the above-describedembodiments in all possible variations thereof is encompassed by theinvention unless otherwise indicated herein or otherwise clearlycontradicted by context.

Groupings of alternative embodiments, elements, or steps of the presentinvention are not to be construed as limitations. Each group member maybe referred to and claimed individually or in any combination with othergroup members disclosed herein. It is anticipated that one or moremembers of a group may be included in, or deleted from, a group forreasons of convenience and/or patentability. When any such inclusion ordeletion occurs, the specification is deemed to contain the group asmodified thus fulfilling the written description of all Markush groupsused in the appended claims.

Unless otherwise indicated, all numbers expressing a characteristic,item, quantity, parameter, property, term, and so forth used in thepresent specification and claims are to be understood as being modifiedin all instances by the term “about.” As used herein, the term “about”means that the characteristic, item, quantity, parameter, property, orterm so qualified encompasses a range of plus or minus ten percent aboveand below the value of the stated characteristic, item, quantity,parameter, property, or term. Accordingly, unless indicated to thecontrary, the numerical parameters set forth in the specification andattached claims are approximations that may vary. At the very least, andnot as an attempt to limit the application of the doctrine ofequivalents to the scope of the claims, each numerical indication shouldat least be construed in light of the number of reported significantdigits and by applying ordinary rounding techniques. Notwithstandingthat the numerical ranges and values setting forth the broad scope ofthe invention are approximations, the numerical ranges and values setforth in the specific examples are reported as precisely as possible.Any numerical range or value, however, inherently contains certainerrors necessarily resulting from the standard deviation found in theirrespective testing measurements. Recitation of numerical ranges ofvalues herein is merely intended to serve as a shorthand method ofreferring individually to each separate numerical value falling withinthe range. Unless otherwise indicated herein, each individual value of anumerical range is incorporated into the present specification as if itwere individually recited herein.

The terms “a,” “an,” “the” and similar referents used in the context ofdescribing the present invention (especially in the context of thefollowing claims) are to be construed to cover both the singular and theplural, unless otherwise indicated herein or clearly contradicted bycontext. All methods described herein can be performed in any suitableorder unless otherwise indicated herein or otherwise clearlycontradicted by context. The use of any and all examples, or exemplarylanguage (e.g., “such as”) provided herein is intended merely to betterilluminate the present invention and does not pose a limitation on thescope of the invention otherwise claimed. No language in the presentspecification should be construed as indicating any non-claimed elementessential to the practice of the invention.

Specific embodiments disclosed herein may be further limited in theclaims using “consisting of” or “consisting essentially of” language.When used in the claims, whether as filed or added per amendment, thetransition term “consisting of” excludes any element, step, oringredient not specified in the claims. The transition term “consistingessentially of” limits the scope of a claim to the specified materialsor steps and those that do not materially affect the basic and novelcharacteristic(s). Embodiments of the present invention so claimed areinherently or expressly described and enabled herein.

All patents, patent publications, and other publications referenced andidentified in the present specification are individually and expresslyincorporated herein by reference in their entirety for the purpose ofdescribing and disclosing, for example, the compositions andmethodologies described in such publications that might be used inconnection with the present invention. These publications are providedsolely for their disclosure prior to the filing date of the presentapplication. Nothing in this regard should be construed as an admissionthat the inventors are not entitled to antedate such disclosure byvirtue of prior invention or for any other reason. All statements as tothe date or representation as to the contents of these documents isbased on the information available to the applicants and does notconstitute any admission as to the correctness of the dates or contentsof these documents.

While aspects of the invention have been described with reference to atleast one exemplary embodiment, it is to be clearly understood by thoseskilled in the art that the invention is not limited thereto. Rather,the scope of the invention is to be interpreted only in conjunction withthe appended claims and it is made clear, here, that the inventorbelieves that the claimed subject matter is the invention.

What is claimed is:
 1. A microorganism evaluation system comprising aviewing section for image acquisition, the viewing section comprising: aviewing section body inlet and an opposite viewing section body outletand a viewing port therebetween, the viewing port configured toaccommodate a fluid flow therethrough from the viewing section bodyinlet to and out of the viewing section body outlet; imaging equipmentoperably installed on the viewing section adjacent to the viewing port;at least one independently controlled imaging light source operablyinstalled in the viewing section and configured to selectivelyilluminate the viewing port, the viewing section being configured incooperation with the imaging equipment and the at least one imaginglight source such that multiple image acquisition events relating to thefluid flow and a discrete microorganism therein are undertaken via theimaging equipment for analysis; and at least one independentlycontrolled light stimulation device operably installed in the viewingsection and configured to selectively emit visible light, the at leastone light stimulation device being configured as a wavelength stimulatorfor selectively emitting narrow band light energy, wherein: a firstillumination event is associated with operation of the at least oneimaging light source; a second illumination event is associated withoperation of the at least one light stimulation device; and the firstand second illumination events and at least one image acquisition eventoccur within a single cycle of the system defined as the time periodfrom the start of one discrete first illumination event of the at leastone imaging light source to the next, whereby the system allows forcontinuous viewing section flow throughput and synchronizes illuminationof the at least one imaging light source and the at least one lightstimulation device of the viewing section for multiple illuminationevents in the single cycle and improved image acquisition by the imagingequipment.
 2. The system of claim 1 wherein the first and secondillumination events are non-overlapping.
 3. The system of claim 2wherein the second illumination event of the at least one lightstimulation device represents approximately eighty to ninety percent(80-90%) of the cycle.
 4. The system of claim 1 wherein the first andsecond illumination events are overlapping.
 5. The system of claim 4wherein the second illumination event of the at least one lightstimulation device represents approximately eighty to one-hundredpercent (80-100%) of the cycle.
 6. The system of claim 1 wherein thecycle is approximately fifteen to forty-five milliseconds (15-45 ms). 7.The system of claim 1 wherein the viewing section comprises twoindependently controlled light stimulation devices, a first lightstimulation device configured to emit light at a first wavelengthserving to stimulate a first microorganism and a second lightstimulation device configured to emit light at a second wavelengthserving to stimulate a second microorganism.
 8. The system of claim 7wherein the first wavelength is approximately 530 nm (green) serving asan attractant to a first microorganism.
 9. The system of claim 8 whereinthe first light stimulation device has a domed, semi-spherical lens fordiffusing light.
 10. The system of claim 7 wherein the second wavelengthis approximately 475 nm (blue) serving to trigger photosynthesis in asecond microorganism.
 11. The system of claim 1 wherein the at least oneimaging light source is installed in the viewing section so as toprovide substantially side illumination within the viewing port.
 12. Thesystem of claim 11 wherein a plurality of imaging light sources arealigned as an array within the viewing port.
 13. The system of claim 1wherein the at least one light stimulation device is operably installedin the viewing section body outlet.
 14. The system of claim 1 whereinthe at least one imaging light source and the at least one lightstimulation device are co-located.
 15. A method of illuminationsynchronization within a microorganism evaluation system comprising aviewing section for fluid flow therethrough and image acquisition, themethod comprising: flowing a fluid through a viewing port from a viewingsection body inlet to and out of a viewing section body outlet, imagingequipment being operably installed on the viewing section adjacent tothe viewing port; independently controlling an at least one imaginglight source operably installed in the viewing section to selectivelyilluminate the viewing port, the viewing section being configured incooperation with the imaging equipment and the at least one imaginglight source such that multiple image acquisition events relating to thefluid flow and a discrete microorganism therein are undertaken via theimaging equipment for analysis; independently controlling an at leastone light stimulation device operably installed in the viewing sectionto selectively emit visible light, the at least one light stimulationdevice being configured as a wavelength stimulator for selectivelyemitting narrow band light energy, a first illumination event beingassociated with operation of the at least one imaging light source, anda second illumination event being associated with operation of the atleast one light stimulation device, the first and second illuminationevents and at least one image acquisition event occurring within asingle cycle of the system defined as the time period from the start ofone discrete first illumination event of the at least one imaging lightsource to the next; and acquiring an image of a microorganism during thesingle cycle, whereby use of the system allows for continuous viewingsection flow throughput and synchronizing illumination of the at leastone imaging light source and the at least one light stimulation deviceof the viewing section for multiple illumination events in the singlecycle and improved image acquisition.
 16. The method of claim 15 whereinthe step of independently controlling an at least one imaging lightsource defines an imaging light source first illumination eventrepresenting approximately five to fifteen percent (5-15%) of the cycle.17. The method of claim 16 wherein the step of independently controllingan at least one light stimulation device defines a light stimulationdevice second illumination event representing approximately eighty toone-hundred percent (80-100%) of the cycle.
 18. The method of claim 15wherein the cycle is approximately fifteen to forty-five milliseconds(15-45 ms).
 19. The method of claim 15 wherein the first and secondillumination events are substantially non-overlapping.
 20. The method ofclaim 15 wherein the step of independently controlling an at least onelight stimulation device comprises independently controlling a firstlight stimulation device configured to emit light at a first wavelengthserving to stimulate a first microorganism and independently controllinga second light stimulation device configured to emit light at a secondwavelength serving to stimulate a second microorganism.